The HIV Interaction in Viral Evolution (HIVE) Center came together a little more than four years ago to understand, at the atomic, biophysical and evolutionary level, the system interdependency of interacting HIV macromolecules and their assemblies which shape the HIV life cycle. To accomplish this goal the proposed program will continue to explore the structural and biophysical interactions of HIV Gag and Gag-Pol polyproteins, capsid, reverse transcriptase, and integrase and their evolutionary relationships. The Center's research focus will also extend to cellular factors that inform the structural and macromolecular dynamics of events in reverse transcription, assembly, and integration. This will include studies on how APOBEC3 proteins suppress reverse transcription, and the role of LEDGF in guiding viral integration and its contribution to latency in conjunction with CPSF6. Studies on HIV drug resistance will tie together a genetic and structural perspective, based on mutational correlations that are due to constraints on protein structural stability and function and which ultimately shape fitness. The depth of the computational strength of the program is centered on expanding structural, biophysical, and viral sequencing findings for developing predictive structural models and small molecule probes targeting viral function. The strength of the Center's biological and computational research will provide insights into the interrelationships of viral and host mechanisms, enabling discovery of new drug targets and therapeutic strategies that may ultimately lead to a cure. The HIVE Center comprises a group of highly collaborative investigators with deep experience in HIV research and well established expertise in structural, biophysical, biochemical, and computational biology, as well as synthetic chemistry, and virology. They will study the mechanistic implications of viral and viral-host macromolecular interactions along with the dynamics and the impacts of the evolution of drug resistance to address relevant biological questions with the following Specific Aims: AIM1: Defining HIV polyproteins and their components in retroviral assembly and maturation AIM 2: Determining HIV-host interactions driving virus reverse transcription and integration AIM 3: Understanding evolution of antiviral resistance mutations and their biological and biophysical implications through studies AIM 4: Developing and characterizing novel small molecule probes to understand biological function